本文選題：聲動(dòng)力療法　切入點(diǎn)：低強(qiáng)度超聲　出處：《重慶醫(yī)科大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 背景:鼻咽癌(Nasopharyngeal carcinoma)是一種好發(fā)于中國南方的惡性腫瘤。目前的治療方法存在一定的局限性和嚴(yán)重的副作用。尋找選擇性好、副作用小的有效方法一直是人們追求的目標(biāo)。 光動(dòng)力治療(photodynamic therapy, PDT)是一種選擇性好、副作用相對小的腫瘤治療方法。它主要是利用選擇性潴留于增殖活躍組織的光敏劑在一定波長的光照下產(chǎn)生細(xì)胞毒作用的活性氧物質(zhì)摧毀腫瘤組織細(xì)胞。由于光在生物組織中的穿透能力有限,嚴(yán)重影響到光動(dòng)力治療的實(shí)際應(yīng)用。令人欣慰的是,有研究發(fā)現(xiàn)一些光敏劑也能被超聲活化。這一發(fā)現(xiàn)激起了人們對聲動(dòng)力治療(Sonodynamic therapy, SDT)研究的興趣。相繼發(fā)現(xiàn)卟啉類光敏劑極易被超聲激活產(chǎn)生活性氧物質(zhì),并得到廣泛深入的研究。竹紅菌素B(hypocrellin B, HB)是新近開發(fā)成功的一種中草藥光敏劑,其成分單一,化學(xué)結(jié)構(gòu)明確,性質(zhì)穩(wěn)定,是一種很有發(fā)展前景的新型光敏劑。開展竹紅菌素B成為一種新型聲敏劑引起我們的廣泛興趣,因此,本研究采用超聲頻率為1.7 MHz.聲強(qiáng)為0.65 W/cm2為實(shí)驗(yàn)參數(shù),探討了低強(qiáng)度超聲聯(lián)合竹紅菌素B對鼻咽癌CNE2細(xì)胞株的影響,旨在為聲動(dòng)力治療提供新的聲敏劑,并為鼻咽癌的臨床治療提供新的技術(shù)手段。 材料與方法:采用倒置顯微鏡觀察和CCK-8法檢測低強(qiáng)度超聲活化HB聲動(dòng)力作用后CNE2細(xì)胞的生長狀況;應(yīng)用流式細(xì)胞檢測術(shù)和Hoechst33258細(xì)胞核熒光染色法觀察超聲活化HB介導(dǎo)的聲動(dòng)力作用對CNE2細(xì)胞凋亡的影響;通過Rhodamine123染色法了解細(xì)胞線粒體膜電位變化情況;最后通過透射電鏡觀察聲動(dòng)力作用后細(xì)胞超微結(jié)構(gòu)的變化情況。 結(jié)果:HB能顯著增強(qiáng)低強(qiáng)度超聲對人鼻咽癌CNE2細(xì)胞的殺傷效率,CCK-8法檢測發(fā)現(xiàn)隨著時(shí)間的增加細(xì)胞抑制率較對照組顯著增加,倒置顯微鏡下觀察到HB介導(dǎo)的聲動(dòng)力組細(xì)胞數(shù)量較其他對照組明顯減少,細(xì)胞固縮,單純HB對細(xì)胞沒有細(xì)胞毒性作用;Hoechst33258細(xì)胞核熒光染色示US(0.65 W/cm2)活化HB(2.5uM)介導(dǎo)的聲動(dòng)力組出現(xiàn)核凝聚、凋亡小體;同時(shí)流式細(xì)胞術(shù)檢測細(xì)胞凋亡率發(fā)現(xiàn)HB介導(dǎo)的聲動(dòng)力組,在聲動(dòng)力作用18小時(shí)候細(xì)胞早期凋亡率達(dá)18.61%; Rhodamine123染色對CNE2細(xì)胞線粒體膜電位檢測顯示HB介導(dǎo)的聲動(dòng)力組,在聲動(dòng)力作用4小時(shí)后細(xì)胞線粒體膜電位明顯降低,最后透射電鏡觀察到HB介導(dǎo)的聲動(dòng)力組,在聲動(dòng)力作用18小時(shí)細(xì)胞出現(xiàn)線粒體腫脹溶解,大部分細(xì)胞壞死,碎裂。 結(jié)論:低強(qiáng)度超聲活化HB介導(dǎo)的聲動(dòng)力作用能夠有效的殺傷人鼻咽癌CNE2細(xì)胞,提示HB可能是一種新型聲敏劑,具有潛在的推廣和應(yīng)用價(jià)值;SDT有望發(fā)展成為一種很有潛力的治療腫瘤的新模式。
[Abstract]:Background: nasopharyngeal carcinoma (NPC) is a malignant tumor in southern China. There are some limitations and serious side effects in the treatment of nasopharyngeal carcinoma. Photodynamic therapy (PDT) is a good selectivity. Cancer treatment with relatively small side effects. It mainly uses reactive oxygen species (Ros), which selectively trap active proliferating tissues to produce cytotoxic effects under certain wavelengths of light. Because of light, tumor cells are destroyed. With limited penetration in biological tissues, Has had a serious impact on the practical use of photodynamic therapy. Some studies have found that some Guang Min agents can also be activated by ultrasound. This discovery has aroused people's interest in the study of sonodynamic therapy (SDTs). It is a newly developed Chinese herbal medicine Guang Min. Its composition is single, its chemical structure is clear, and its properties are stable. It is a new type of Guang Min agent with great development prospects. It is of great interest to develop a new type of acoustical sensitizer. Therefore, the ultrasonic frequency is 1.7 MHz and the sound intensity is 0.65 W / cm ~ 2 as the experimental parameter in this study, and the experimental parameters are as follows: (1) the ultrasonic frequency is 1.7 MHz and the sound intensity is 0.65 W / cm ~ (2). The effects of low intensity ultrasound combined with erythromycin B on nasopharyngeal carcinoma (NPC) CNE2 cell line were studied in order to provide a new sound sensitizer for acoustic dynamic therapy and a new technique for clinical treatment of nasopharyngeal carcinoma. Materials and methods: the growth of CNE2 cells was observed by inverted microscope and CCK-8 method. Flow cytometry and Hoechst33258 nuclear fluorescence staining were used to observe the effect of ultrasonic activated HB-mediated acoustics on apoptosis of CNE2 cells, and the changes of mitochondrial membrane potential were observed by Rhodamine123 staining. At last, the ultrastructure of the cells was observed by transmission electron microscope (TEM). Results the effect of low intensity ultrasound on human nasopharyngeal carcinoma (NPC) CNE2 cells was significantly enhanced by 1: HB. The results of CCK-8 method showed that the inhibition rate of human nasopharyngeal carcinoma (NPC) CNE2 cells was significantly higher than that of the control group with the increase of time. Under inverted microscope, the number of cells in HB-mediated acoustic power group was significantly lower than that in other control groups, and cell pyknosis was observed in HB-mediated acoustical power group. There was no cytotoxic effect of HB alone on cells. Fluorescence staining of Hoechst33258 cells showed that US(0.65 W / cm 2) activated HB2. 5 uM) -mediated nuclear condensation and apoptotic corpuscles were found in the acoustic power group, and the apoptosis rate was detected by flow cytometry in the HB-mediated acoustic dynamic group, and the cell apoptosis rate was found in the HB-mediated acoustics group by flow cytometry. The rate of early apoptosis reached 18.61% at 18 hours after acoustics, and the mitochondrial membrane potential of CNE2 cells was significantly decreased by Rhodamine123 staining after 4 hours of acoustical stimulation, and the mitochondrial membrane potential of CNE2 cells was detected by Rhodamine123 staining. Finally, transmission electron microscopy showed that in HB-mediated acoustic power group, mitochondria swelling and dissolution, necrosis and fragmentation of most cells were observed at 18 hours after acoustical stimulation. Conclusion: low intensity ultrasound activated HB-mediated acoustic action can effectively kill human nasopharyngeal carcinoma (NPC) CNE2 cells, suggesting that HB may be a new type of sound sensitizer. SDT is expected to become a potential new model for tumor treatment.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R739.63

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本文編號(hào)：1595662